Association of 410L, 1016I and 1534C kdr mutations with pyrethroid resistance in Aedes aegypti from Ouagadougou, Burkina Faso, and development of a one-step multiplex PCR method for the simultaneous detection of 1534C and 1016I kdr mutations

Abstract

Background Since 2000, Burkina Faso has experienced regular dengue cases and outbreaks, making dengue an increasingly important health concern for the country. Previous studies in Burkina Faso reported that resistance of Aedes aegypti to pyrethroid insecticides was associated with the F1534C and V1016I kdr mutations. The current study reports high resistance of Ae. aegypti populations to pyrethroid insecticides, likely supported by mutations in the voltage-gated sodium channel, here evidenced by genotyping the kdr SNPs V410L, V1016I and F1534C. We also describe a new multiplex PCR-based diagnostic of F1534C and V1016I kdr SNPs. Methods Larvae of Ae. aegypti were collected from three health districts of Ouagadougou in 2018. The resistance status of Ae. aegypti to permethrin (15 μg/ml) and deltamethrin (10 μg/ml) was tested using bottles and to malathion (5%) using WHO tube tests. All bioassays used 1-h exposure and mortality recorded 24 h post-exposure. Bioassay results were interpreted according to WHO thresholds for resistance diagnosis. The kdr mutations were screened using AS-PCR and TaqMan methods in exposed and non-exposed Aedes mosquitoes. Results Females from all health districts were resistant to permethrin and deltamethrin (< 20% mortality) but were fully susceptible to 5% malathion. The F1534C and V1016I kdr mutations were successfully detected using a newly developed multiplex PCR in perfect agreement with TaqMan method. The 1534C/1016I/410L haplotype was correlated with permethrin resistance but not with deltamethrin resistance; however, the test power was limited by a low frequency of dead individuals in deltamethrin exposure. Conclusions Resistance to pyrethroid insecticides is associated with kdr mutant haplotypes, while the absence of substantial resistance to malathion suggests that it remains a viable option for dengue vector control in Ouagadougou.

First report of V1016I, F1534C and V410L kdr mutations associated with pyrethroid resistance in Aedes aegypti populations from Niamey, Niger

“…The V1016I and V410L mutations appear linked in Aedes populations from Niger. Similar trend has been found in Latin America and in Burkina Faso [ 22 , 42 ]. However, in Ghana [ 25 ] the dissociation appears much greater suggesting a different stage of co-evolution for the two mutants [ 42 ].…”

“…We also found the CIL haplotype to be associated with both permethrin and deltamethrin resistance. In contrast, in Ouagadougou it was associated only with permethrin resistance [ 22 ].…”

“…The diversity of tri-locus genotypes (10) and tri-locus haplotypes (6) that we found suggest a late emergence of the insecticide resistance compared to Ouagadougou in Burkina Faso where the 1534C mutation is almost fixed, effectively reducing the number of genotypes (6) and haplotypes (3) and to the three mutations [ 22 ].…”

“…Detection of the V410L kdr mutation was performed following the protocol of Granada et al . [ 35 ]with modifications in the PCR conditions described in Sombié et al [ 22 ].…”

“…In West Africa, two kdr mutations, V1016I and F1534C have been reported in Cape Verde [ 20 ] whereas in Nigeria, S989P and F1534C kdr mutations are reported [ 21 ]. Three kdr mutations, V410L, V1016I and F1534C have been reported from Burkina Faso [ 22 , 23 ], Côte d’Ivoire [ 24 ] and Ghana [ 25 ].…”